Objectives: This study aims to determine if the CNTNAP2 gene polymorphism at the rs7794745 locus is associated with autism spectrumdisorder (ASD) by comparing its frequency in autistic children and healthy controls. Methods: The case-control study involved a total of 150 samples, comprising 80 individuals diagnosed with autism spectrum disorder(ASD) and 70 healthy children. These participants, both male and female, had an average age of 3.5 ± 3 years for autistic individuals and4.2 ± 3 years for healthy children. Genomic analysis was performed by amplifying and scrutinizing DNA sequences through the utilizationof polymerase chain reaction (PCR) and restriction enzymes. Specifically, the genotyping of the CNTNAP2 gene at the rs7794745 locus wasaccomplished using the PCR-RFLP method. To facilitate genotyping, the isolation of DNA from peripheral blood cells sourced from healthychildren and patients. Results: Our findings indicated a statistically non-significant low-frequency distribution (P-value > 0.05) of the rs7794745 SNP in ASDpatients compared to healthy children. Conclusion: The study’s findings propose that there is no correlation between the rs7794745 polymorphism and autism spectrum disorder (ASD).